Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters fat – fatty oil – ester-type wax – or...
Patent
1995-01-27
1998-08-11
Campell, Bruce R.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters fat, fatty oil, ester-type wax, or...
800250, 800255, 800DIG17, 800DIG69, 800DIG70, 4351723, 4353201, 536 241, 935 6, 935 35, 935 64, C12N 500, C12N 1500, C07H 2104
Patent
active
057929220
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to upstream DNA sequences and their use to control expression of genes in developing plant seeds and their use.
BACKGROUND
Studies in plant gene expression have yielded a number of general conclusions concerning the elements that control expression. Plants, like other organisms both prokaryotic and eukaryotic, contain conserved or consensus sequences upstream (5') of the transcriptional start site of genes which appear capable of regulating transcriptional rates. In eukaryotes, these sequences include a motif found typically about 25 bp 5' to the transcriptional initiation site which has the sequence TATAA/TAA/T and is referred to as a TATA box. The role of this TATA box appears to be to define the transcriptional start for RNA polymerase II. A second upstream sequence is referred to as a CAAT box. Typically, this is found about 75 bases upstream of the transcriptional start and is associated with regulating the frequency of transcriptional initiation. In plants the consensus sequence may be either CCAAT or sometimes AGGA. However, neither of these alternative consensus sequences need be present in all plant genes. These sequence motifs and their DNA context within 70-90 bases upstream of the transcriptional start are often referred to as promoters. In general, 5' of the promoter region and most frequently within 2000 bases of it are cis-acting elements which confer a variety of properties on the promoter and which can modulate transcriptional activity in either a constitutive or a non-constitutive manner. These cis-acting sequences may be referred to as enhancers (if they are responsible for increases in transcription) or silencers (if they are responsible for decreases or suppression of transcription). Enhancers and silencers are frequently the sites at which nuclear proteins bind or interact. The modulating nuclear proteins are called trans-acting factors. They are considered to be very important for non-constitutive or regulated expression as they may be the major determinant of the activity of a gene in a particular tissue or organ or in response to an external stimulus. The relationship between this protein binding and the enhancer/silencer element may determine the transcriptional activity. The isolation of genes which are activated by heat, light or chemicals such as endogenous hormones or are activated in specific organs such as seeds, leaves or flowers has permitted analysis of factors which may determine how expression is regulated. In numerous, but not all, cases, it has been shown that the construction of chimeric genes which contain the promoter and optionally cis-elements from a given regulated gene and a coding sequence of a reporter protein not normally associated with that promoter gives rise to regulated expression of the reporter. The use of promoters from seed-specific genes for the expression of sequences in seed of genes that are either not normally expressed in a seed-specific manner or those that require an altered pattern of expression has been attempted on only a few occasions. In all cases to date, chimeric genes designed for seed-specific expression have used seed-storage protein regulatory signals and promoters. However, it is evident from work on storage protein gene expression that expression commences at a fairly late stage in embryogenesis, namely once the embryo has reached (in the case of dicots) the classical torpedo shape. Thus, although storage proteins express at high levels and their regulation is often transcriptional, the timing and level of expression may not be ideal for all seed-specific applications. It is, therefore, of interest to identify other seed-specific promoters and enhancers with temporal or cellular specificity different from that of seed storage proteins, such as those from oleosins.
The following disclose organ or tissue-specific regulatory sequences used to produce tissue- or organ-specific expression in transformed plants. There are several by now "classical" examples of regulated gene expression in non
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Campell Bruce R.
Sembiosys Genetics Inc.
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